Chromatography: Monosaccharides
- Chromatography is a technique that can be used to separate a mixture into its individual components
- Chromatography relies on differences in the solubility of the different chemicals (called ‘solutes’) within a mixture
- All chromatography techniques use two phases:
- The mobile phase
- The stationary phase
- The components in the mixture separate as the mobile phase travels over the stationary phase
- Differences in the solubility of each component in the mobile phase which affects how far each component can travel
- Those components with higher solubility will travel further than the others
- This is because they spend more time in the mobile phase and are thus carried further up the paper than the less soluble components
Paper Chromatography
- Paper chromatography is one specific form of chromatography
- In paper chromatography:
- The mobile phase is the solvent in which the sample molecules can move, which in paper chromatography is a liquid e.g. water or ethanol
- The stationary phase in paper chromatography is the chromatography paper
Paper chromatography method
- A spot of the mixture (that you want to separate) is placed on chromatography paper and left to dry
- The chromatography paper is then suspended in a solvent
- As the solvent travels up through the chromatography paper, the different components within the mixture begin to move up the paper at different speeds
- Larger molecules move slower than smaller ones
- This causes the original mixture to separate out into different spots or bands on the chromatography paper
- This produces what is known as a chromatogram
An example of a chromatogram that has been produced by using paper chromatography to separate a spot of ink
Using chromatography to separate a mixture of Monosaccharides
- Paper chromatography can be used to separate a mixture of monosaccharides
- Mixtures containing coloured molecules, such as ink or chlorophyll, do not have to be stained as they are already coloured
- Mixtures of colourless molecules, such as a mixture of monosaccharides, have to be stained first
- A spot of the stained monosaccharide sample mixture is placed on a line at the bottom of the chromatography paper
- Spots of known standard solutions of different monosaccharides are then placed on the line beside the sample spot
- The chromatography paper is then suspended in a solvent
- As the solvent travels up through the chromatography paper, the different monosaccharides within the mixture separate out at different distances from the line
- The unknown monosaccharides can then be identified by comparing and matching them with the chromatograms of the known standard solutions of different monosaccharides
- If a spot from the monosaccharide sample mixture is at the same distance from the line as a spot from one of the known standard solutions, then the mixture must contain this monosaccharide
How chromatography can be used to separate a mixture of monosaccharides and identify the individual components
Examiner Tip
Paper chromatography is the name given to the overall separation technique while a chromatogram is the name given to the visual output of a chromatography run. This is the piece of chromatography paper with the visibly separated components after the run has finished.